Diversifying interferon functions through combinatorial and structural biology

通过组合和结构生物学使干扰素功能多样化

• 批准号: 8633201
• 负责人: Juan Luis Mendoza
• 金额: $ 11.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-18 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8633201
• 关键词: Affinity; Basic Science; Binding; Biochemistry; Biological; Biological Assay; Biology; Cancer Biology; Cell membrane; Cells; Complex; Computational Biology; Computer Simulation; Coupled; Critiques; Cytokine Receptors; Cytokine Signaling; DNA; Development; Dimerization; Disease; Engineering; Equilibrium; Event; Evolution; Exhibits; Family; Flowcharts; Future; Goals; Health; Human; Immune; Immune System Diseases; Immune response; In Vitro; Individual; Interferon Receptor; Interferon Type I; Interferons; Ligand Binding; Ligands; Malignant Neoplasms; Mediating; Mentors; Methodology; Molecular; Molecular Structure; Nature; Output; Physiological Processes; Play; Property; Publishing; Receptor Signaling; Relative (related person); Research Proposals; Role; Scientist; Shapes; Signal Pathway; Signal Transduction; Specificity; Structure; Structure-Activity Relationship; System; Techniques; Testing; Therapeutic; Toxic effect; Training; Translating; Variant; Viral; Virus; Virus Diseases; Vision; Work; Writing; arm; base; cancer cell; cancer therapy; cell growth; combinatorial; cytokine; design; directed evolution; extracellular; improved; insight; meetings; member; molecular recognition; novel; programs; public health relevance; receptor; receptor binding; receptor-mediated signaling; research and development; research study; response; structural biology; therapeutic development; tool; tumor

PROJECT SUMMARY/ABSTRACT: In this KO1 proposal, we attempt to deeply interrogate the role of molecular recognition and structure in a receptor-ligand system, type I Interferons (IFNs), that has fundamental implications for basic receptor signaling biology as well as human health. We intend to elucidate the structure- functional relationships of IFN induced signaling and to enhance IFN functions through combinatorial, computational, and structural biology. The training and research proposal describes a five year mentored program under the guidance of Dr. K. Christopher Garcia designed to build upon Dr. Mendoza's previous training in order to further expertise in combinatorial, computational, and structural biology. This will establish Dr. Mendoza as an independent multi-disciplinary scientist with a powerful set of tools to give him the experimental range to tackle problems in cytokine engineering for cancer biology. We will attempt to understand how IFN cytokines mediate a wide range of physiological processes through their effects on cell growth, differentiation, and proliferation. Type I IFNs have anti-proliferative properties important for the surveillance and control of malignant cells. Upon engagement of their receptor extracellular regions, IFNs differentially activate intracellular JAK/STAT signaling cascades. Members of the type I IFN family elicit different signaling responses despite binding to a common pair of receptors, IFN¿R1 and IFN¿R2. Fundamentally, the specificity of cytokine signaling is largely determined by the composition of receptor chains within the signaling complex. While much is known about cytokine-activated intracellular signaling pathways, we know much less about how extracellular structural changes induced by ligand binding translates into signaling events. The experiments proposed will provide more clarity to this question. The potential translational and cancer therapeutic applications are based on a pioneering study published more than ten years ago suggesting that IFN diversification through in vitro evolution could yield molecules with enhanced functional properties. Now, armed with powerful new methodologies, we wish to revisit this concept to create IFNs with diverse functional properties, but also to provide a structural and mechanistic rationale for these altered activities in a way that could inform strategies to engineer IFNs for cancer therapy in the future. We will achieve this by 1) in vitro evolution of IFN¿1, 2) identification of variants with altered function using in vitro anti-viral and anti-proliferative assays, 3) structural characterization of the ternary complexes of the evolved IFNs to correlate structure with function, and 4) the computational design and in vitro evolution of chimeric cytokines with distinct and novel signaling and functional outputs. By understanding the molecular basis of cytokine/receptor interactions with correlated function, the results from this work may be able to guide the future development of cytokines as therapeutics as well as expand the capabilities and diversity of receptor mediated signaling, functional activities, and therapeutic properties.

项目摘要/摘要：在此KO1提案中，我们试图深入询问 接收器配体系统中的分子识别和结构，I型干扰素（IFNS），具有基本的 对基本受体信号生物学以及人类健康的影响。我们打算阐明结构 IFN诱导信号传导的功能关系，并通过组合增强IFN功能， 计算和结构生物学。培训和研究建议描述了五年任期 在K. Christopher Garcia博士的指导下进行的计划，旨在建立在Mendoza博士之前 培训以进一步的组合，计算和结构生物学专业知识。这将建立 门多萨博士是一名独立的多学科科学家，拥有一套强大的工具，可以给他 解决癌症生物学细胞因子工程问题的实验范围。 我们将尝试了解IFN细胞因子如何介导广泛的物理过程 通过它们对细胞生长，分化和增殖的影响。 I型IFN具有抗增殖 对监视和控制恶性细胞重要的特性。接收器订婚 细胞外区域，IFN不同地激活细胞内JAK/Stat信号级联。成员 I型IFN家族引起不同的信号响应目的地与普通的接收器，IFN¿R1和 ifn？r2。从根本上讲，细胞因子信号的特异性在很大程度上取决于 信号复合物中的受体链。虽然对细胞因子激活的细胞内知识已有很多了解 信号通路，我们对配体结合引起的细胞外结构变化的了解少得多 转化为信号事件。提出的实验将为这个问题提供更明确的清晰度。这 潜在的转化和癌症治疗应用是基于一项开创性的研究 超过十年前，提示通过体外进化多样化可以产生分子 增强功能性能。现在，拥有强大的新方法，我们希望重新审视这个概念 创建具有不同功能属性的IFN，但也提供结构和机械原理 这些改变的活动以一种可以告知策略将来为癌症治疗设计策略的方式。 我们将通过1）IFN¿的体外演变1，2）识别具有改变功能的变体 使用体外抗病毒和抗增殖测定，3）三元复合物的结构表征 进化的IFN将结构与函数相关联，以及4）计算设计和体外演变 具有独特和新颖的信号传导和功能输出的嵌合细胞因子。通过了解分子 细胞因子/受体相互作用与相关功能的基础，这项工作的结果可能能够指导 细胞因子作为治疗的未来发展以及扩大接收器的能力和多样性 介导的信号传导，功能活动和治疗特性。